Supplement: Metalloendocrinology: Mapping Bioinorganic Chemistry in the Extracellular Space

项目摘要 铜是生物学中必需的微量营养素，在呼吸和呼吸等过程中发挥着重要作用。 抗氧化防御铜离子所处的环境对其活性至关重要， 错误划分和失衡与许多疾病有关。虽然技术 出现了研究细胞内铜动力学，细胞外铜池相对不足， 表征了这一补充的母基金旨在阐明细胞外金属的形态 空间，重点是金属与细胞外环境中另一种动态成分的相互作用， 荷尔蒙本课题的主要研究对象是一种具有类肽功能的多肽，即C- 肽，衍生自胰岛素原蛋白的31聚体。历史上被低估为一个无辜的次要标志 对于胰岛素，C肽由于其不依赖于胰岛素的生物活性而越来越受到关注。然而，它的进步 由于缺乏对机械的理解而受到阻碍。在父项目下的工作表明，C- 肽可以直接与铜结合，这是一种可以调节C肽活性的事件。在本建议的目标1中，我们 试图确定C肽及其截短物是否以及如何影响铜的运输。前期工作 表明金属结合相互作用可以改变金属的稳态平衡。一个结合在 体外和体内工作将试图分析驱动C肽和铜之间相互作用的途径。 目的2试图利用铜和C肽的相互作用来鉴定质膜受体， 肽。虽然已经提出了一种孤儿GPCR，GPR146，作为C肽的伴侣，但这种说法仍然存在。 辩论。我们假设金属与富含负电荷残基的肽的相互作用， 可能影响受体识别。这一目标将筛选与孤儿库的C肽相互作用 在存在和不存在过渡金属的情况下的GPCR，以确定金属是否是受体的组成部分 识别，并且如果是，则鉴定肽的同源受体。为了实现这些研究目标，候选人 将通过多方面的指导团队进行跨学科的培训， 金属内分泌过程的分子洞察力。

PROJECT ABSTRACT Copper is an essential micronutrient in biology, playing fundamental roles in processes such as respiration and antioxidant defense. The context in which a copper ion resides is vital to its activity, and its miscompartmentalization and imbalances is associated with a host of disorders. While technologies have emerged to investigate intracellular copper dynamics, the extracellular copper pool is relatively under- characterized. The parent grant of this supplement seeks to elucidate the speciation of metals in the extracellular space, with a focus on the interplay of metals with another dynamic component in the extracellular environment, hormones. The main research of this objective focuses on a particular peptide with hormone-like functions, C- peptide, a 31-mer derived from the proinsulin protein. Historically undervalued as an innocent secondary marker for insulin, C-peptide is of increasing interest for its bioactivity independent of insulin. However, its advancement has been hampred by lack of mechanistic understanding. Work under the parent project has shown that C- peptide can directly bind to copper, an event that can modulate C-peptide's activity. In Aim 1 of this proposal, we seek to determine whether and how C-peptide and its truncates affect copper trafficking. Preliminary work suggests that the metal-binding interaction can shift the homeostatic balance of the metal. A combination of in vitro and in vivo work will seek to analyze the pathways that drive the interplay between C-peptide and copper. Aim 2 seeks to leverage the interactions of copper and C-peptide to identify a plasma membrane receptor for the peptide. While an orphan GPCR, GPR146, has been proposed as a partner for C-peptide, this claim remains debated. We hypothesize that metal interactions with the peptide, which is rich in negatively-charged residues, may be influential in receptor recognition. This aim will screen for C-peptide interactions with a library of orphan GPCRs in the presence and absence of transition metals to determine whether metals are integral to receptor recognition, and if so, identify a cognate receptor for the peptide. To achieve these research goals, the candidate to be supported by this supplement will be trained across disciplines with a multifaceted mentoring team to bring molecular insight to metalloendocrinological processes.